Transcription mechanisms that contribute to HIV-1 latency

导致 HIV-1 潜伏期的转录机制

基本信息

批准号：
8468555
负责人：
Andrew J Henderson
金额：
$ 41.25万
依托单位：
BOSTON MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8468555
关键词：
Anti-Retroviral Agents Architecture Biochemical Biochemical Process CD4 Positive T Lymphocytes Cell Line Cells Chromatin Complex Coupled Data Event Exhibits Genes Genetic Transcription Goals HIV HIV Infections HIV Long Terminal Repeat HIV-1 In Vitro Infection Interruption Libraries Life Long Terminal Repeats Maintenance Mass Spectrum Analysis Molecular Phenotype Play Positive Transcriptional Elongation Factor B Properdin Proteomics Proviruses RNA Polymerase II Recruitment Activity Regulation Repression Role Source System T memory cell T-Lymphocyte Tissues Transcription Elongation Transcription Initiation Transcription Repressor/Corepressor Viral Proteins Virus cellular targeting chromatin immunoprecipitation chromatin remodeling gene repression insight macrophage negative elongation factor novel polarized cell premature promoter public health relevance purge success termination factor transcription termination

项目摘要

DESCRIPTION (provided by applicant): HIV provirus is regulated at the level of transcription and involves changes in transcription initiation, chromatin organization and elongation. In particular, transcription elongation has been demonstrated to be a limiting step for HIV expression. We hypothesize that the coordinated control of RNA polymerase II processivity and premature transcription termination coupled with chromatin remodeling creates a key checkpoint that limits provirus transcription and directly impact HIV latency in different cellular and molecular contexts. We have shown that RNA polymerase II processivity is a check point for HIV transcription and that NELF, a factor that pauses RNA polymerase II, re-enforces repression of HIV proviruses by recruiting a termination factor Pcf11 and transcriptional corepressors to the HIV-1 long terminal repeat. Using a variety of biochemical approaches, including chromatin immunoprecipitation, RNAPII footprinting, in vitro transcription systems and mass spectrometry, we propose to determine the biochemical mechanisms and characterize the cellular factors that negatively regulate HIV transcription elongation. These studies will provide new insights into the establishment, maintenance and reversal of HIV latency. Understanding the regulation of HIV transcription elongation will provide novel cellular targets for controlling and purging HIV in different cellular reservoirs.

描述（由申请人提供）：HIV病毒在转录水平上受到调节，涉及转录启动，染色质组织和伸长率的变化。特别是，已证明转录伸长是HIV表达的限制步骤。我们假设对RNA聚合酶II加工性的协调控制和与染色质重塑相结合的过早转录终止会产生一个关键检查点，该检查点限制了Profirus转录并直接影响不同细胞和分子环境中的HIV潜伏期。我们已经表明，RNA聚合酶II加工性是HIV转录的一个检查点，而NELF（暂停RNA聚合酶II，通过募集终止因子PCF11和转录核心pcf11和对HIV-1长时间重复的终止因子PCF11和转录核心pc）来增强对HIV Provirus的抑制。使用多种生化方法，包括染色质免疫沉淀，RNAPII足迹，体外转录系统和质谱法，我们建议确定生化机制，并表征对HIV转录延长的负面调节的细胞因素。这些研究将为艾滋病毒潜伏期的建立，维持和逆转提供新的见解。了解HIV转录伸长的调节将为控制和清除不同细胞储层中的HIV提供新颖的细胞靶标。